Cell Division Differentiation Research Articles

Differential cell division history between neutrophils and macrophages in their development from granulocyte–macrophage progenitors

The appearance of monocytes before neutrophils in the blood during haematopoietic recovery in myelosuppressive patients is commonly observed, thus suggesting a difference in the cell division history between these two lineages in the differentiation from granulocyte-macrophage (GM) progenitors. We investigated the cell division histories of murine GM progenitors. When analysed by the dye dilution method, GM progenitors gave rise to Gr-1+Fms+ and Gr-1+Fms- cells that passed through similar rounds of cell division during initial 5 d of culture. The Gr-1+Fms+ cells showed morphological features of monocytes, while Gr-1+Fms- cells exhibited an immature morphology of neutrophils. In the subsequent culture, a decline in the number of Gr-1+Fms+ cells was observed, while Gr-1+Fms- cells increased. The proliferation of Gr-1+Fms- cells and no cell division of Gr-1+Fms+ cells were confirmed by DNA staining, Ki-67 expression, membrane dye staining and bromodeoxyuridine incorporation. These Gr-1+Fms- cells acquired mature neutrophil morphology, whereas Gr-1+Fms+ cells became macrophages. These results demonstrate that GM progenitors generate postmitotic monocytes earlier than mature neutrophils. Our data may also offer one explanation for the rapid recovery of monocytes in comparison with neutrophils in the early phase of haematopoietic regeneration.

Cellular and Subcellular Structure of Neoproterozoic Animal Embryos

Stereoblastic embryos from the Doushantuo Formation of China exhibit occasional asynchronous cell division, with diminishing blastomere volume as cleavage proceeded. Asynchronous cell division is common in modern embryos, implying that sophisticated mechanisms for differential cell division timing and embryonic cell lineage differentiation evolved before 551 million years ago. Subcellular structures akin to organelles, coated yolk granules, or lipid vesicles occur in these embryos. Paired reniform structures within embryo cells may represent fossil evidence of cells about to undergo division. Embryos exhibit no evidence of epithelial organization, even in embryos composed of approximately 1000 cells. Many of these features are compatible with metazoans, but the absence of epithelialization is consistent only with a stem-metazoan affinity for Doushantuo embryos.

Auxin, Ethylene and Brassinosteroids: Tripartite Control of Growth in the Arabidopsis Hypocotyl

Dark-grown Arabidopsis seedlings develop an apical hook by differential cell elongation and division, a process driven by cross-talk between multiple hormones. Auxins, ethylene and gibberellins interact in the formation of the apical hook. In the light, a similar complexity of hormonal regulation has been revealed at the level of hypocotyl elongation. Here, we describe the involvement of brassinosteroids (BRs) in auxin- and ethylene-controlled processes in the hypocotyls of both light- and dark-grown seedlings. We show that BR biosynthesis is necessary for the formation of an exaggerated apical hook and that either application of BRs or disruption of BR synthesis alters auxin response, presumably by affecting auxin transport, eventually resulting in the disappearance of the apical hook. Furthermore, we demonstrate that ethylene-stimulated hypocotyl elongation in the light is largely controlled by the same mechanisms as those governing formation of the apical hook in darkness. However, in the light, BRs appear to compensate for the insensitivity to ethylene in hls mutants, supporting a downstream action of BRs. Hence, our results indicate that HLS1, SUR1/HLS3/RTY1/ALF1 and AMP1/HPT/COP2/HLS2/PT act on the auxin-ethylene interaction, rather than at the level of BRs. A model for the tripartite hormone interactions is presented.

Society for Reproductive Biology Founders' Lecture 2003. The making of an embryo: short-term goals and long-term implications.

During early development, the eutherian mammalian embryo forms a blastocyst comprising an outer trophectoderm epithelium and enclosed inner cell mass (ICM). The short-term goal of blastocyst morphogenesis, including epithelial differentiation and segregation of the ICM, is mainly regulated autonomously and comprises a combination of temporally controlled gene expression, cell polarisation, differentiative cell divisions and cell-cell interactions. This aspect of blastocyst biogenesis is reviewed, focusing, in particular, on the maturation and role of cell adhesion systems. Early embryos are also sensitive to their environment, which can affect their developmental potential in diverse ways and may lead to long-term consequences relating to fetal or postnatal growth and physiology. Some current concepts of embryo-environment interactions, which may impact on future health, are also reviewed.

Society for Reproductive Biology Founders' Lecture 2003.The making of an embryo: short-term goals and long-term implications.

During early development, the eutherian mammalian embryo forms a blastocyst comprising an outer trophectoderm epithelium and enclosed inner cell mass (ICM). The short-term goal of blastocyst morphogenesis, including epithelial differentiation and segregation of the ICM, is mainly regulated autonomously and comprises a combination of temporally controlled gene expression, cell polarisation, differentiative cell divisions and cell–cell interactions. This aspect of blastocyst biogenesis is reviewed, focusing, in particular, on the maturation and role of cell adhesion systems. Early embryos are also sensitive to their environment, which can affect their developmental potential in diverse ways and may lead to long-term consequences relating to fetal or postnatal growth and physiology. Some current concepts of embryo–environment interactions, which may impact on future health, are also reviewed.

Actual targets in cytodifferentiation cancer therapy.

Transformation of a normal cell into a tumor cell results from six essential alterations in cell physiology. There is a complex relationship that exists between growth, differentiation, neoplastic transformation, and the expression of genes and tumor suppressor genes. The knowledge of these mechanisms demonstrates that it is possible to pharmacologically modulate the growth and differentiation of tumor cells. The differentiation therapy focuses on demonstrating that cancer is a reversible state with altered maturation in which the transformed phenotype may be suppressed by cytostatic agents and by the pharmacological differentiation towards benign forms with no proliferative potential. One of the mechanisms determining the activity of target genes is the post-translational modification of the N-terminal tails of core histones. Inappropriate repression of genes required for cell differentiation has been linked to several forms of cancer. Histone deacetylase inhibitors modulate transcription, and are endowed with cytodifferentiating, antiproliferative and apoptogenic properties. Retinoids modulate cell differentiation, proliferation, apoptosis and morphogenesis in vertebrates, and have proved to be clinically useful. Their biological effects are mediated by the activation of retinoic acid receptors, which are ligand-dependent gene transcription factors. Checkpoints during cell cycle allow the cell to respond to proliferation signals or decide between the alternate pathways leading to cytokinesis, differentiation, quiescence, and cell death. Abrogation of normal cell cycle controls in tumor cells contributes to their inability to differentiate and the restoration of such controls in G1 can lead to the resumption of differentiation and terminal cell division. Chemical inhibitors of cyclin-dependent kinases have been reported to stimulate differentiation of tumor-cell lines.

Early patterning of the mouse embryo--contributions of sperm and egg.

The first cleavage of the fertilised mouse egg divides the zygote into two cells that have a tendency to follow distinguishable fates. One divides first and contributes its progeny predominantly to the embryonic part of the blastocyst, while the other, later dividing cell, contributes mainly to the abembryonic part. We have previously observed that both the plane of this first cleavage and the subsequent order of blastomere division tend to correlate with the position of the fertilisation cone that forms after sperm entry. But does sperm entry contribute to assigning the distinguishable fates to the first two blastomeres or is their fate an intrinsic property of the egg itself? To answer this question we examined the distribution of the progeny of early blastomeres in embryos never penetrated by sperm - parthenogenetic embryos. In contrast to fertilised eggs, we found there is no tendency for the first two parthenogenetic blastomeres to follow different fates. This outcome is independent of whether parthenogenetic eggs are haploid or diploid. Also unlike fertilised eggs, the first 2-cell blastomere to divide in parthenogenetic embryo does not necessarily contribute more cells to the blastocyst. However, even when descendants of the first dividing blastomere do predominate, they show no strong predisposition to occupy the embryonic part. Thus blastomere fate does not appear to be decided by differential cell division alone. Finally, when the cortical cytoplasm at the site of sperm entry is removed, the first cleavage plane no longer tends to divide the embryo into embryonic and abembryonic parts. Together these results indicate that in normal development fertilisation contributes to setting up embryonic patterning, alongside the role of the egg.

C-Jun modulates apoptosis but not terminal cell differentiation in murine erythroleukemia cells.

The proto-oncogene c-Jun has been implicated in the control of cell proliferation and differentiation and more recently in the regulation of apoptosis. We have previously reported the involvement of c-Jun in the erythroid differentiation block in murine erythroleukemia (MEL) cells. As reported here, we investigated the role of c-Jun in the regulation of terminal differentiation and apoptosis of MEL cells by studying different stable transfectant clones containing c-jun constructs in sense or antisense orientation. c-Jun did not prevent cell growth arrest in G0/G1 and p21 induction that are normally associated with terminal differentiation induced by DMSO treatment, suggesting that c-Jun may uncouple phenotypic differentiation and terminal cell division in the MEL cell system. Spontaneous apoptosis was accelerated in c-jun expressing MEL cells before and after DMSO treatment. Moreover, c-Jun sensitized apoptosis induced by various drugs. Drug-induced apoptosis was associated with c-Jun N-terminal kinase (JNK) activation and c-Jun N-terminal phosphorylation (JNP). In contrast, overexpression of c-jun delayed apoptosis in serum-starved cells, indicating that c-Jun may reduce or accelerate apoptosis in MEL cells depending on the nature of the apoptotic stimulus. These results suggest that the proto-oncogene c-Jun may modulate differentiation and apoptosis of leukemic cells.

Reprogramming leukemic cells to terminal differentiation by inhibiting specific cyclin-dependent kinases in G1.

Some tumor cells can be stimulated to differentiate and undergo terminal cell division and loss of tumorigenicity. The in vitro differentiation of murine erythroleukemia (MEL) cells is a dramatic example of tumor-cell reprogramming. We found that reentry of MEL cells into terminal differentiation is accompanied by an early transient decline in the activity of cyclin-dependant kinase (CDK) 2, followed by a decline of CDK6. Later, as cells undergo terminal arrest, CDK2 and CDK4 activities decline. By analyzing stable MEL-cell transfectants containing vectors directing inducible expression of specific CDK inhibitors, we show that only inhibitors that block the combination of CDK2 and CDK6 trigger differentiation. Inhibiting CDK2 and CDK4 does not cause differentiation. Importantly, we also show that reprogramming through inhibition of CDKs is restricted to G(1) phase of the cell cycle. The results imply that abrogation of normal cell-cycle controls in tumor cells contributes to their inability to differentiate fully and that restoration of such controls in G(1) can lead to resumption of differentiation and terminal cell division. The results also indicate that CDK4 and CDK6 are functionally distinct and support our hypothesis that the two CDKs regulate cell division at different stages of erythroid maturation.

Role of cell division in branching morphogenesis and differentiation of the embryonic pancreas.

A new culture system for the embryonic pancreas enables the formation of a branched organ in vitro. In such cultures, each terminal branch originates as a small bud and the number of buds and of terminal branches increases progressively with the expansion of the culture. However buds can also be resorbed during growth. The normal labelling index of cells in incipient buds ("tips") is greater than between buds ("dips") suggesting that budding may be driven by a local increase of cell division. Consistent with this, treatments that reduce cell division repress the formation of buds and branches. It is not possible to initiate budding in isolated endodermal epithelium by treatment with fibroblast growth factor, although this does increase the degree of differentiation of exocrine cells. Cultures in which cell division is completely inhibited by aphidicolin treatment will produce more endocrine cells than usual and inhibit the differentiation of exocrine cells. Consistent with this it is found that in untreated cultures the division of endocrine precursors cannot be detected by BrdU labelling whereas the division of exocrine precursors is frequent. It is concluded that cell division is necessary for bud formation in the embryonic pancreas and that the growth factors required for this normally come from the mesenchyme. Cell division is also necessary for exocrine differentiation. Endocrine cells, however, can arise from undifferentiated progenitors without cell division.

Reprogramming erythroleuekmia cells to terminal differentiation and terminal cell division

Reprogramming erythroleuekmia cells to terminal differentiation and terminal cell division

Reprogramming erythroleuekmia cells to terminal differentiation and terminal cell division

In vitro differentiation of murine erythroleuekemia cells recapitulates many aspects of the erythroid terminal differentiation program, including hemoglobin synthesis and proliferation arrest. It also provides an opportunity to study the changes occurring during reprogramming of tumor cells into their normal differentiation program. This review is focused on the recent progress made in understanding the key events occurring during the reprogramming of erythroleukemia cells. We discuss the contributions of PU.1 to the block to terminal differentiation exhibited by the erythroleukemia cells as well as the role of GATA-1 in restoring normal differentiation. We also discuss the role of certain cell cycle regulators in the decision to resume normal differentiation and in the resulting terminal cell divisions and arrest.

Kinetics of T cell turnover following thymectomy.

We develop a mathematical formula that provides the number of cells in an isolated population that have divided k times in n days (O< or =k< or =n). This differential cell division formula is applied to the kinetics of peripheral T cells in the diabetes-prone BB rat following thymectomy. These rats received daily intraperitoneal injections of the DNA precursor, bromodeoxyuridine (BrdUrd), over a period of 12-13 days. As the cells divided, they incorporated BrdUrd progressively into their DNA molecules, and the differential formula provides a close prediction of the fraction of BrdUrd-positive T cells present each day during this 'pulse' phase. No further BrdUrd was administered after 13 days, and the diminishing fraction of BrdUrd-positive cells was recorded for several more weeks. The differential cell division formula was capable of describing the rather complex form of the retention curve as BrdUrd-tagged DNA molecules passed to progeny cells during this 'chase' phase. We believe that this simple formula may be found generally useful in describing cell kinetic data in mitotically active cells.

Determination and development of the larval muscle pattern in Drosophila melanogaster.

This review describes briefly what is known about the early steps of mesoderm differentiation in the fruitfly Drosophila melanogaster. After a summary of general aspects including mesoderm differentiation, mesoderm cell migration and subdivision of the mesoderm, more detail is given about the specification of muscle progenitor cells, due to their role as the earliest obvious landmarks in muscle fiber development in Drosophila. Particular focus is given to recent results on the role of asymmetric cell division in muscle differentiation. Furthermore a short summary of myoblast fusion is provided.

Asymmetric cell division and cell fate in plants

A variety of approaches has recently been employed to investigate how sister cells adopt distinct fates following asymmetric divisions during plant development. Surgical and drug studies have been used to analyze asymmetric divisions during both early embryogenesis in brown algae and pollen development in tobacco. Genetic screens have been used to identify genes in Arabidopsis thaliana that are required for specific asymmetric cell divisions during pollen and root development. These studies indicate that cell polarity and division orientation are closely tied to the process of cell fate specification, and suggest that differential inheritance of determinants and positional information may both be involved in the specification of cell fates following asymmetric cell division.

Deregulated expression of the PU.1 transcription factor blocks murine erythroleukemia cell terminal differentiation.

Murine erythroleukemia (MEL) cells are transformed erythroid precursors that are blocked from completing the late stages of erythroid differentiation. A frequent event in the generation of these malignant cells is deregulation of the hematopoietic-specific transcription factor PU.1 (Spi-1) by retroviral insertion of the spleen-focus-forming virus component of Friend virus. During chemically induced reinitiation of MEL cell terminal differentiation, expression of PU.1 is rapidly down-regulated, suggesting that PU.1 might interfere with processes required for terminal differentiation of erythroid precursors. To investigate the role of PU.1 in erythroid differentiation we transfected MEL cells with a PU.1 cDNA controlled by the eucaryotic translation elongation factor EF1 alpha promoter. Deregulated expression of PU.1 blocked chemically induced differentiation and terminal cell division. Deregulated expression of two other protooncogenes, c-myc and c-myb, also has been shown to block MEL differentiation. We present evidence that PU.1 inhibits terminal differentiation at an earlier step than c-Myc and c-Myb. Thus reinitiation of MEL cell terminal differentiation appears to be controlled by an ordered program of turning off several protooncogenes. Down-regulation of PU.1 may be a very early step in this program.

Organization and cell differentiation in lateral roots of Arabidopsis thaliana.

Lateral root formation in plants involves the stimulation of mature pericycle cells to proliferate and redifferentiate to create a new organ. The simple organization of the root of Arabidopsis thaliana allows the development of lateral root primordia to be characterized histologically. We have divided the process of lateral root development into 8 stages defined by specific anatomical characteristics and cell divisions. To identify the cell types in the developing primordium we have generated a collection of marker lines that express beta-glucuronidase in a tissue- or cell type-specific manner in the root. Using these tools we have constructed a model describing the lineage of each cell type in the lateral root. These studies show that organization and cell differentiation in the lateral root primordia precede the appearance of a lateral root meristem, with differential gene expression apparent after the first set of divisions of the pericycle.

Pollinium development and number in the Orchidaceae

Development of anthers in three subfamilies of Orchidaceae was studied anatomically to examine homology hypotheses for pollinium number characters and to produce a model of pollinium development for the family. Serial sections of plastic‐embedded embryonic inflorescences revealed that anther primordia were either flattened or ovoid; subsequent expansion of thecae and their inward (adaxial) reorientation (“rotation”), achieved by differential cell division and elongation in the connective, result in a mature anther with strongly introrse morphology and pollinia oriented side by side (juxtaposed). Strongly introrse anthers occur in at least some members of all subfamilies and are probably the basal state for the family. All anthers examined (from Orchidoideae, Spiranthoideae, and Epidendroideae) showed a single meristematic region, which would later give rise to pollen, per theca at earliest stages; septation of each of these regions resulted in four or eight pollinia per anther, while lack of septation in some members of the Epidendroideae gave two pollinia. In contrast, the two bipartite pollinia found in many Spiranthoideae and Orchidoideae were produced by adherence of the contents of two locules at a late ontogenetic stage, and should be recognized as a distinct character state. Eight pollinia result from partitioning by two longitudinal septa or a longitudinal and a transverse septum; these two morphologies may also represent separate character states.

A New Class of Arabidopsis Constitutive Photomorphogenic Genes Involved in Regulating Cotyledon Development.

Light signals have profound effects on morphogenesis of hypocotyls and cotyledons of Arabidopsis seedlings, but the mechanisms by which light signals are transduced and integrated to control these processes are poorly understood. We report here the identification of a new class of constitutive photomorphogenic (cop) mutants, cop2, cop3, and cop4, in which dark-grown seedlings have open and enlarged cotyledons resembling those of light-grown wild-type seedlings. The epistatic relationships of these three mutations to previously characterized phytochrome-deficient mutations suggest that COP2, COP3, and COP4 may act downstream of phytochrome in the light regulatory pathway. Mutations in each of the three loci alleviate the normal inhibition of cell-type differentiation, cell enlargement, and lateral cell division observed in cotyledons of dark-grown wild-type seedlings, but do not affect plastid differentiation. The cop4 mutation also leads to high-level dark expression of nuclear, but not plastid-encoded, light-inducible genes. We further show that for the nuclear cab1 gene encoding a chlorophyll a/b binding protein of the photosynthetic light-harvesting complex, activation in dark-grown cop4 mutants is achieved by modulation of promoter activity. Interestingly, COP4 modulates cab1 promoter activity through a pathway distinct from that of COP1 and COP9. Furthermore, cop4 mutants are defective in both root and shoot gravitropic responses, indicating that the COP4 locus may be involved in both light-signaling and gravity-sensing processes.

REGULATION OF THE CELL DIVISION CYCLE AND DIFFERENTIATION IN BACTERIA

INTRODUCTION AND OBJECTIVES . . . . . . . .. . . . . ... .. . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . .. 690 THE CAULOB'ACTER LIFE CYCLE ... 691 Asymmetric Cell Division . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 691 Chromosome Replication and Segregation . . . . . . . . . . . . . . . . . 692 Sequence of Developmental Events 694 GENETIC TOOLS AND APPROACHES. . . . . . . . . .. . . . . . . . . . . . ... . . .. . . . . . . . . . . . . .. . . .. . . . . . . . . . 694 CDC GENES AND CELL CYCLE REGULATION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 695 Mutational Analysis 695 Functional Organization of CDC Genes 696 Genetic Mapping 697 MORPHOGENETIC PATHWAYS AND TEMPORAL CONTROL. . . . . . . . . . . . . . . . . . . . . . 698 Pilus Assembly 698 Flagellum Biosynthesis . . . . . . ...... . . . . . . . . . . . .. . . . . . . . . . 698 Regulatory Pathways in Other Systems . . . .. . . . . . . . . . . . . . .. . . . . . . .. . . . . . . . . . . 702 PROMOTER SPECIFICITY AND TRANSCRIPTION REGULATION . . . . . . . . . . . . . . . . . . . 703 a'4 Promoters 704 Other Promoters 706 Regulatory Proteins and Transcriptional Activation 707 PROTEIN TARGETING AND SPATIAL LOCALIZATION ... 708 Flagellar Proteins 708 Chemotactic Proteins . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 709 Heat-Shock Proteins 710 ROLE OF CDC GENES IN DEVELOPMENTAL REGULATION . . . . . . . . . . . . . . . . . . . . . . . . 7 1 0 Developmental Phenotypes of CDC Mutants 7 1 0 Pseudoreversion Analysis 711 SUMMARY AND PROSPECTS. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . 7 1 3